CN104788527B - Method for extracting tripterine efficiently with ionic liquid - Google Patents

Method for extracting tripterine efficiently with ionic liquid Download PDF

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CN104788527B
CN104788527B CN201510129853.XA CN201510129853A CN104788527B CN 104788527 B CN104788527 B CN 104788527B CN 201510129853 A CN201510129853 A CN 201510129853A CN 104788527 B CN104788527 B CN 104788527B
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tripterine
extraction
water
hsccc
volume ratio
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CN104788527A (en
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蔡美强
苏洁
金米聪
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Zhejiang Gongshang University
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    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

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Abstract

The invention relates to a method for extracting tripterine efficiently with an ionic liquid. The method comprises the following steps: 1) ultrasonic enhanced extraction: an extracting solution is a mixed solution of ethanol, water and an imidazolium ionic liquid, wherein the volume ratio of ethanol to water to imidazolium ionic liquid is 1: (0.01-0.5):(0.5-2.0), and ultrasonic treatment is performed; 2) organic solvent extraction: the extraction is performed according to the volume ratio of the extracting solution to water to C1-C3 haloalkanes to be (1-5):(1-3):1, and an organic phase is subjected to concentration under reduced pressure to be approximately dried to obtain a tripterine extract; (3) column chromatography purification: the tripterine extract and silica gel are fully mixed and then the mixture is packed, and the packed mixture is subjected to petroleum ether leaching and elution and volatilization to dryness with a petroleum ether-ethyl acetate mixed solvent to obtain a crude tripterine product; 4) HSCCC separation and purification; (5) vacuum freeze drying. The method has the advantages that the application of the organic solvent is reduced greatly, the pollution and harm in the extraction process are reduced remarkably, and the extraction effect and the extraction efficiency are improved.

Description

A kind of method of utilization ionic liquid high efficiency extraction tripterine
Technical field
The present invention relates to technology of pharmaceutical engineering field, more particularly to it is a kind of using ionic liquid high efficiency extraction tripterine Method.
Background technology
It is Celastraceae tripterygium plant that Radix Tripterygii Wilfordii is called Caulis Fibraureae (Tripterygium wilfordii Hook.f.), is faced It is widely used in the treatment autoimmune disease such as rheumatoid arthritiss on bed, its instant effect, determined curative effect, almost not having can be with The similar Chinese medicine for substituting completely.Tripterine is the native chemical with higher physiologically active produced in Radix Tripterygii Wilfordii plant Thing, because which has special chemical constitution and mechanism of action, particularly clinically to autoimmune disease (such as erythema wolf Skin ulcer, rheumatoid arthritiss, ankylosing spondylitises and anaphylactoid purpura etc.) there is unique curative effect, it is anti-that pharmacological research proves which has Inflammation, immunosuppressant, antitumor and anti-AIDS etc. are acted on.Therefore, carry out effective ingredient in Radix Tripterygii Wilfordii (such as tripterine) Purification technique is studied the research and development to following new drug and has far-reaching influence.Tripterine structural formula is as follows：
At present, extract tripterine from the plant material containing tripterine mainly to enter using volatile organic solvent Row is extracted, and combines high-speed countercurrent chromatography (HSCCC) using liquid/liquid extraction binding silica gel column chromatography or liquid/liquid extraction.Such as patent The Chinese invention patent of Application No. 201210230388.5 (Publication No. 103524591A) be " a kind of prepared from active ingredients of tripterygium wilfordii Preparation method " a kind of method that the separation from Radix et Rhizoma Tripterygii prepares triptolide there is provided silica gel column chromatography.The patent with Radix et Rhizoma Tripterygii is raw material, and the double-aqueous phase system formed using polymerinorganic saline solution and ethanol is extracted, then insoluble with water Property organic solvent back extraction, then in conjunction with the refined tripterine of HSCCC methods.Number of patent application is 201310574275.1 (public Accuse number be 103570795A) Chinese invention patent《A kind of preparation method of tripterine》, the patent of invention is by fresh south Mabi root drying and crushing, is soaked using hexane solution, and the normal hexane extractum of acquisition obtains thunder Jing after the purification of column chromatography for separation twice Celastrol.Chinese invention patent of the number of patent application for 201310027337.7 (notification number is 103087142A)《One kind is carried The method for taking tripterine》, the plant material containing tripterine adopts methanol or ethanol to soak, obtained by the patent of invention The tripterine alcohol extract of 0.005% (W/W) is not less than to content, then uses C1～C5Halogenated alkane extraction, and by extract Tripterine is obtained after concentrating and using silica gel post separation.The tripterine purity that these methods are obtained can reach 95% with On, but substantial amounts of organic solvent has been used in extraction and in separating preparation, the safety and pollution that there is organic solvent is asked Topic.
The special performance of ionic liquid is widely paid close attention in chemical field, is separated in the extraction of natural product at present In succeed application, the such as middle promulgated by the State Council of Patent No. ZL200810114416.0 (Authorization Notice No. is 101597296 B of CN) Bright patent《A kind of new method of efficiently extracting and producing artemisinin by ionic liquid》Using glyoxaline ion liquid halide salts and water Mixture it is ultrasound-enhanced extract Herba Artemisiae Annuae raw material in arteannuin.At present with regard to using ionic liquid from Radix et Rhizoma Tripterygii, stem, Ye Zhong The method for extracting tripterine has not been reported, therefore, carry out the ionic liquid extract research of tripterine, to solving to work as It is front to extract that to separate a large amount of potential safety hazards caused using organic reagent in preparation process significant.
The content of the invention
The technical problem to be solved is provided one kind and safely, effectively utilizes ionic liquid for prior art The method that body extracts tripterine.
The present invention solve the technical scheme that adopted of above-mentioned technical problem for：It is a kind of to utilize ionic liquid high efficiency extraction Thunder God The method of rattan red pigment, it is characterised in that comprise the following steps：
1) ultrasound-enhanced extraction：Plant material drying containing tripterine crosses 40～80 mesh sieves after crushing, by solid-liquid Than 1：5～100 add volume ratio to be 1：0.01～0.5：0.5～2.0 ethanol, I mixed liquor of water and glyoxaline ion liquid, in Ultrasonic power 10～200W/cm of density2And under 10～100kHz of supersonic frequency, supersound extraction 5 under the conditions of 20～65 DEG C～ 90min, must contain the extracting solution of tripterine after solid-liquid separation；
2) organic solvent extraction：By C1～C3Halogenated alkane and a certain amount of water be added to the step 1) carrying of obtaining Take in liquid, by extracting solution, water and C1～C3Halogenated alkane volume ratio 1～5：1～3：1, extract 1～5 time.Separate organic faciess Tripterine extraction phase is obtained, then the organic extraction for obtaining is evaporated at 40～60 DEG C and is closely done, contained The red extractum of tripterine.Extraction extraction raffinate is recycled, and the extraction extraction raffinate Main Ingredients and Appearance is above-mentioned glyoxaline ion liquid Ⅰ；
3) column chromatography purification：By the red extractum containing tripterine for obtaining with silica gel (200～300 mesh) by weight Than 1：10～100 are sufficiently mixed, and are then charged into during a bottom has the glass chromatography column of sieve plate, are first used appropriate petroleum ether Drip washing, then be 4 with volume ratio：1 petroleum ether and ethyl acetate mixed solvent eluting, the flow velocity for controlling eluent are not more than 10mL/min, collects red eluent, then the red eluent for obtaining is evaporated at 40～60 DEG C and is closely done, obtained Tripterine crude product；
4) HSCCC is isolated and purified：HSCCC dicyandiamide solutions are selected, by volume/weight ratio 0.2～2：1 (mL/mg) is by HSCCC Dicyandiamide solution and above-mentioned tripterine crude product are added in separatory funnel, concussion, stratification, after balance, by upper and lower phase point Open, with the upper as fixing phase of dicyandiamide solution, the lower of dicyandiamide solution is mutually mobile phase, adjusts rotating speed, pumps into stream with certain flow rate Dynamic phase, according to the spectrogram Fraction collection object of detector, obtains the solution of tripterine respectively；
5) vacuum lyophilization：By above-mentioned each tripterine solution, under proper temperature, drying pressure, vacuum freezing is done It is dry, organic extractant is reclaimed, tripterine of the purity more than 95% is respectively obtained.
Preferably, the glyoxaline ion liquid I is 1- ethyl-3-methylimidazole villaumites, 1- ethyl-3-methylimidazoles In bromide, 1- butyl -3- Methylimidazole. bromides, 1- octyl group -3- methylimidazole villaumites, 1- pi-allyl -3- methylimidazole villaumites It is at least one.
Preferably, the step 1) in ethanol, I mixed liquor of water and glyoxaline ion liquid volume ratio be 1：0.05： 1。
Preferably, it is 2 that the HSCCC dicyandiamide solutions are volume ratio：3：3：2: 0.05 normal hexane-ethyl acetate-first Alcohol-HSCCC the dicyandiamide solutions of the preparation of water-ionic liquid II, volume ratio are 2：3：3：4: 0.05 methyl tertiary butyl ether(MTBE)-positive fourth The HSCCC dicyandiamide solutions or volume ratio that alcohol-alcohol-water-ionic liquid II is prepared is 1：2：2：1: 0.05 normal hexane-acetic acid second Any one in the HSCCC dicyandiamide solutions that ester-alcohol-water-ionic liquid II is prepared.
Preferably, the glyoxaline ion liquid II is 1- ethyl-3-methylimidazole tetrafluoroborates, 1- ethyl -3- In Methylimidazole. hexafluorophosphate, 1- butyl -3- methyl imidazolium tetrafluoroborates, 1- pi-allyl -3- Methylimidazole. bromides extremely Few one kind.
Compared with prior art, it is an advantage of the current invention that：The present invention is using the ethanol, water for mixing by a certain percentage With I mixed liquor of glyoxaline ion liquid as extracting solution, extract in the plant material containing tripterine with reference to ultrasonic wave added Tripterine, the extracting solution greatlys save the usage amount of organic solvent compared with existing pure organic solvent extraction, green Color, environmental protection, significantly reduce pollution and danger that traditional volatile organic solvent extracting method is brought to environment and experimenter Evil, while the addition of ionic liquid, substantially improves extraction effect, improves extraction efficiency.Further, with ion in the present invention Liquid is used as HSCCC Mobile Phase Additives, hence it is evident that shorten the disengaging time of HSCCC, substantially increases and effect is separated with impurity Rate, the purity of the tripterine for obtaining are more than 95%, and the safety and friendly to environment and human body has more advantage.
Description of the drawings
Ultraviolet chromatograms of the Fig. 1 for the tripterine of the HSCCC in the embodiment of the present invention 1, wherein, (1) is Radix Tripterygii Wilfordii Red pigment.
Specific embodiment
The present invention is described in further detail below in conjunction with accompanying drawing embodiment.
Embodiment 1：
1) ultrasound-enhanced extraction：Radix et Rhizoma Tripterygii 1.0kg is taken, 80 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：10 (W/V) add ethanol/water/1- ethyl-3-methylimidazoles villaumite (10：0.5：8, V/V/V mixture), in ultrasonic power density 100W/cm2, supersonic frequency 100kHz, under the conditions of 40 DEG C of temperature, supersound extraction 30min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Dichloromethane and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, By the volume ratio 3 of extracting solution, water and dichloromethane：2：1 extraction 3 times, separates organic faciess and obtains tripterine extraction phase, then will The organic extraction for obtaining is evaporated at 55 DEG C closely to be done, and obtains the red extractum containing tripterine.Extraction extraction raffinate Recycle, the main component of the extraction extraction raffinate is ionic liquid I (1- ethyl-3-methylimidazole villaumites).
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 100mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 150mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 55 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select normal hexane-acetic acid second Ester-alcohol-water -1- pi-allyl -3- Methylimidazole .s bromide (2：3：3：2: 0.05, V/V) as HSCCC dicyandiamide solutions, by 250mL HSCCC dicyandiamide solutions and above-mentioned tripterine crude product add separatory funnel in, concussion be sufficiently mixed solution, placed At night, after split-phase balance, separate upper and lower phase.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, adjusts rotating speed and is 700r/min, column temperature are 30 DEG C, with the high-speed counter-current chromatograph (TBE-1000A of Shanghai Tongtian Biotechnology Co., Ltd.'s production Type) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, tripterine is obtained Solution.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -20 DEG C of pre-freezing temperature, operating pressure 25Pa, distillation temperature 50 DEG C of degree, 70 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.
Embodiment 2：
1) ultrasound-enhanced extraction：Thunder God rattan 1.0kg is taken, 60 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：30 (W/V) add ethanol/water/1- ethyl-3-methylimidazoles bromide (10：0.2：7, V/V/V mixture), in ultrasonic power density 50W/cm2, supersonic frequency 50kHz, under the conditions of temperature 45 C, supersound extraction 30min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Chloroform and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, The volume ratio of extracting solution, water and chloroform is 3：2：1, extract 3 times, separate organic faciess and obtain tripterine extraction phase, then The organic extraction for obtaining is evaporated at 55 DEG C and is closely done, obtain the red extractum containing tripterine.Extraction is remaining Liquid is recycled, and the extraction extraction raffinate is 1- ethyl-3-methylimidazole bromides.
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 50mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 150mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 45 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select methyl tertiary butyl ether(MTBE)- N-butyl alcohol-acetonitrile-water -1- ethyl-3-methylimidazoles hexafluorophosphate (2：3：3：4: 0.05, v/v) as HSCCC solvent bodies System, the HSCCC dicyandiamide solutions of 250mL and above-mentioned tripterine crude product are added in separatory funnel, and concussion makes solution abundant Mixing, stands overnight, and separates upper and lower phase after split-phase balance.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, Adjustment rotating speed is 750r/min, and column temperature is 30 DEG C, and with high-speed counter-current chromatograph, (Shanghai Tongtian Biotechnology Co., Ltd. produces TBE-1000A types) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, thunder is obtained The solution of celastrol.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -15 DEG C of pre-freezing temperature, operating pressure 30Pa, distillation temperature 50 DEG C of degree, 60 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.
Embodiment 3：
1) ultrasound-enhanced extraction：Radix et Rhizoma Tripterygii 1.0kg is taken, 60 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：20 (W/V) add ethanol/water/1- butyl -3- Methylimidazole .s bromide (10：1：8, V/ V/V mixture), in ultrasonic power density 50W/cm2, supersonic frequency 50kHz, under the conditions of 40 DEG C of temperature, supersound extraction 50min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Dichloromethane and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, The volume ratio of extracting solution, water and dichloromethane is 3：2：2, extract 3 times, separate organic faciess and obtain tripterine extraction phase, then The organic extraction for obtaining is evaporated at 45 DEG C and is closely done, obtain the red extractum containing tripterine.Extraction is remaining Liquid is recycled, and the main component of the extraction extraction raffinate is 1- butyl -3- Methylimidazole. bromides.
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 50mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 200mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 45 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select methyl tertiary butyl ether(MTBE)- N-butyl alcohol-alcohol-water -1- butyl -3- methyl imidazolium tetrafluoroborates (2：3：3：4: 0.05, v/v) as HSCCC solvent bodies System, the HSCCC dicyandiamide solutions of 250mL and above-mentioned tripterine crude product are added in separatory funnel, and concussion makes solution abundant Mixing, stands overnight, and separates upper and lower phase after split-phase balance.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, Adjustment rotating speed is 750r/min, and column temperature is 30 DEG C, and with high-speed counter-current chromatograph, (Shanghai Tongtian Biotechnology Co., Ltd. produces TBE-1000A types) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, thunder is obtained The solution of celastrol.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -15 DEG C of pre-freezing temperature, operating pressure 30Pa, distillation temperature 50 DEG C of degree, 60 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.
Embodiment 4：
1) ultrasound-enhanced extraction：Tripterygium leaf 1.0kg is taken, 60 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：10 (W/V) add ethanol/water/1- octyl group -3- methylimidazole villaumites (10：2：7, V/ V/V mixture), in ultrasonic power density 20W/cm2, supersonic frequency 100kHz, under the conditions of temperature 60 C, supersound extraction 50min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Dichloromethane and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, The volume ratio of extracting solution, water and dichloromethane is 5：4：3, extract 3 times, separate organic faciess and obtain tripterine extraction phase, then The organic extraction for obtaining is evaporated at 45 DEG C and is closely done, obtain the red extractum containing tripterine.Extraction is remaining Liquid is recycled, and the main component of the extraction extraction raffinate is 1- octyl group -3- methylimidazole villaumites.
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 100mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 150mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 45 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select normal hexane-acetic acid second Ester-alcohol-water -1- ethyl-3-methylimidazole hexafluoros borate (1：2：2：1: 0.05, v/v) as HSCCC dicyandiamide solutions, will During the HSCCC dicyandiamide solutions of 250mL and above-mentioned tripterine crude product add separatory funnel, concussion is sufficiently mixed solution, Stand overnight, after split-phase balance, separate upper and lower phase.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, is adjusted Rotating speed is 750r/min, and column temperature is 30 DEG C, with the high-speed counter-current chromatograph (TBE- of Shanghai Tongtian Biotechnology Co., Ltd.'s production 1000A types) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, Radix Tripterygii Wilfordii is obtained The solution of red pigment.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -15 DEG C of pre-freezing temperature, operating pressure 25Pa, distillation temperature 50 DEG C of degree, 50 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.
Embodiment 5：
1) ultrasound-enhanced extraction：Thunder God rattan 1.0kg is taken, 60 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：25 (W/V) add ethanol/water/1- pi-allyl -3- methylimidazole villaumites (10：1：6, V/V/V mixture), in ultrasonic power density 100W/cm2, supersonic frequency 100kHz, under the conditions of temperature 45 C, supersound extraction 20min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Dichloromethane and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, The volume ratio of extracting solution, water and dichloromethane is 3：5：3, extract 3 times, separate organic faciess and obtain tripterine extraction phase, then The organic extraction for obtaining is evaporated at 45 DEG C and is closely done, obtain the red extractum containing tripterine.Extraction is remaining Liquid is recycled, and the main component of the extraction extraction raffinate is 1- pi-allyl -3- methylimidazole villaumites.
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 100mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 100mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 45 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select normal hexane-acetic acid second Ester-alcohol-water -1- pi-allyl -3- methylimidazole villaumites (1：2：2：1: 0.05, v/v) as HSCCC dicyandiamide solutions, by 250mL HSCCC dicyandiamide solutions and above-mentioned tripterine crude product add separatory funnel in, concussion be sufficiently mixed solution, placed At night, after split-phase balance, separate upper and lower phase.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, adjusts rotating speed and is 750r/min, column temperature are 30 DEG C, with the high-speed counter-current chromatograph (TBE-1000A of Shanghai Tongtian Biotechnology Co., Ltd.'s production Type) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, tripterine is obtained Solution.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -20 DEG C of pre-freezing temperature, operating pressure 25Pa, distillation temperature 55 DEG C of degree, 65 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.
Embodiment 6：
1) ultrasound-enhanced extraction：Radix et Rhizoma Tripterygii 1.0kg is taken, 80 mesh sieves is crossed after drying and crushing, 100 grams of powder is then taken and is put into In 2500 milliliters of conical flasks, by solid-to-liquid ratio 1：50 (W/V) add ethanol/water/1- butyl -3- Methylimidazole .s bromide (10：1：7, V/ V/V mixture), in ultrasonic power density 80W/cm2, supersonic frequency 100kHz, under the conditions of temperature 45 C, supersound extraction 30min, Jing solid-liquid separation obtains the extracting solution containing tripterine.
2) organic solvent extraction：Chloroform and a certain amount of water are added in the extracting solution that 1) above-mentioned steps obtain, The volume ratio of extracting solution, water and chloroform is 3：4：3, extract 3 times, separate organic faciess and obtain tripterine extraction phase, then The organic extraction for obtaining is evaporated at 45 DEG C and is closely done, obtain the red extractum containing tripterine.Extraction is remaining Liquid is recycled, and the main component of the extraction extraction raffinate is 1- butyl -3- Methylimidazole. bromides.
3) column chromatography purification：By the red extractum 1g containing tripterine for obtaining and 50g silica gel (200～300 mesh) Load after being sufficiently mixed in glass chromatography column of the bottom with sieve plate, first with 100mL petroleum ether drip washing, then with volume ratio be 4：1 petroleum ether and ethyl acetate mixed solvent 100mL eluting, the flow velocity for controlling eluent are not more than 10mL/min, collect red Then the red eluent for obtaining is evaporated at 45 DEG C and is closely done, obtain tripterine crude product by color eluent；
4) HSCCC is isolated and purified：Weigh step 3) the tripterine crude product 500mg that obtains, select normal hexane-acetic acid second Ester-methanol-water -1- pi-allyl -3- methylimidazole villaumites (2：3：3：2: 0.05, v/v) as HSCCC dicyandiamide solutions, by 200mL HSCCC dicyandiamide solutions and above-mentioned tripterine crude product add separatory funnel in, concussion be sufficiently mixed solution, placed At night, after split-phase balance, separate upper and lower phase.With the upper as fixing phase of dicyandiamide solution, lower is mutually mobile phase, adjusts rotating speed and is 750r/min, column temperature are 30 DEG C, with the high-speed counter-current chromatograph (TBE-1000A of Shanghai Tongtian Biotechnology Co., Ltd.'s production Type) HSCCC serpentine pipes are pumped into 5.0mL/min flow velocitys, object is collected according to the spectrogram of detector, tripterine is obtained Solution.
5) vacuum lyophilization：By step 4) collection liquid that obtains, -20 DEG C of pre-freezing temperature, operating pressure 25Pa, distillation temperature 55 DEG C of degree, 65 DEG C of resolution temperature obtain tripterine of the purity more than 95%.
Using Agilent 1100series high performance liquid chromatographs, chromatographic column is Zorbax XDB-C for detection18Post (150mm × 4.6mm I.D., 5 μm, Agilent Technologies, USA), mobile phase is 0.05% acetic acid-ammonium acetate (5mmol/L) solution/methanol (25：75, V/V), flow velocity is 0.6mL/min, and column temperature is 35 DEG C, and UV-detector is in 425nm ripples Detected under length, the purity of tripterine is more than 95%.

Claims (4)

1. a kind of method of utilization ionic liquid high efficiency extraction tripterine, it is characterised in that comprise the following steps：
1) ultrasound-enhanced extraction：Plant material drying containing tripterine crosses 40～80 mesh sieves after crushing, by solid-to-liquid ratio 1：5 ～100 add volume ratio to be 1：0.01～0.5：0.5～2.0 ethanol, I mixed liquor of water and glyoxaline ion liquid, in ultrasound 10～200W/cm of power density2And under 10～100kHz of supersonic frequency, 5～90min of supersound extraction under the conditions of 20～65 DEG C, Gu Liquid must contain the extracting solution of tripterine after separating；
2) organic solvent extraction：By C1～C3Halogenated alkane and a certain amount of water be added to the step 1) extracting solution that obtains In, by extracting solution, water and C1～C3Halogenated alkane volume ratio 1～5：1～3:1 extraction 1～5 time；Separate organic faciess and obtain Thunder God Then the organic extraction for obtaining is evaporated at 40～60 DEG C and is closely done, obtained containing Radix Tripterygii Wilfordii by rattan red pigment extraction phase The red extractum of red pigment.
3) column chromatography purification：By the silica gel of the red extractum containing tripterine for obtaining and 200～300 mesh by weight 1： 10～100 are sufficiently mixed, and are then charged into during a bottom has the glass chromatography column of sieve plate, are first drenched with appropriate petroleum ether Wash, then be 4 with volume ratio：1 petroleum ether and ethyl acetate mixed solvent eluting, the flow velocity for controlling eluent are not more than 10mL/ Min, collects red eluent, then the red eluent for obtaining is evaporated at 40～60 DEG C and is closely done, obtain Thunder God Rattan red pigment crude product；
4) HSCCC is isolated and purified：HSCCC dicyandiamide solutions are selected, by volume/weight ratio 0.2～2：1 (mL/mg) is by HSCCC solvents System and above-mentioned tripterine crude product are added in separatory funnel, concussion, stratification, after balance, are mutually separated upper and lower, With the upper as fixing phase of dicyandiamide solution, the lower of dicyandiamide solution is mutually mobile phase, adjusts rotating speed, pumps into flowing with certain flow rate Phase, according to the spectrogram Fraction collection object of detector, obtains the solution of tripterine respectively；
5) vacuum lyophilization：By above-mentioned each tripterine solution, the vacuum lyophilization under proper temperature, drying pressure, Organic extractant is reclaimed, tripterine of the purity more than 95% is respectively obtained
Wherein, described HSCCC dicyandiamide solutions are：Volume ratio is 2：3：3：2: 0.05 normal hexane-ethyl acetate-alcohol-water- The HSCCC dicyandiamide solutions of 1- pi-allyl -3- Methylimidazole. bromides, volume ratio are 2：3：3：4: 0.05 methyl tertiary butyl ether(MTBE)-just The HSCCC dicyandiamide solutions of butanol-acetonitrile-water -1- ethyl-3-methylimidazole hexafluorophosphates, or volume ratio be 2：3：3：4∶ In the HSCCC dicyandiamide solutions of 0.05 methyl tertiary butyl ether(MTBE)-n-butyl alcohol-alcohol-water -1- butyl -3- methyl imidazolium tetrafluoroborates One kind.
2. as claimed in claim 1 using the method for ionic liquid high efficiency extraction tripterine, it is characterised in that：The miaow Azole ionic liquid I is 1- ethyl-3-methylimidazole villaumites, 1- ethyl-3-methylimidazole bromides, 1- butyl -3- Methylimidazole .s At least one in bromide, 1- octyl group -3- methylimidazole villaumites, 1- pi-allyl -3- methylimidazole villaumites.
3. as claimed in claim 1 using the method for ionic liquid high efficiency extraction tripterine, it is characterised in that：The step It is rapid 1) in ethanol, I mixed liquor of water and glyoxaline ion liquid volume ratio be 1：0.05：1.
4. as claimed in claim 1 using the method for ionic liquid high efficiency extraction tripterine, it is characterised in that：The C1 ～C3Halogenated alkane be chloroform, dichloromethane, 1,2- dichloroethanes, 1,2- dichloroethanes and carbon tetrachloride at least It is a kind of.
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